A METHOD TO ISOLATE CDNA-QUALITY RNA FROM ADULT CONIFER NEEDLES AND A PSBA CDNA FROM NORWAY SPRUCE

摘要

In order to investigate the expression of the psbA gene in damaged and undamaged Norway spruce trees (Picea abies) a cDNA clone encoding the D1, protein was isolated via RT-PCR. Applying a method developed by Schneiderbauer er al. (1991) with some modifications, we were able to obtain the required RNA from mature needles and successfully reverse transcribe it into cDNA. Sequence analysis of the cDNA clone revealed an open reading frame (ORF) encoding a 353 amino acid polypeptide that is highly homologous to the D1 protein sequences deduced from higher plant psbA genes. A 4 bp insertion, directly following the stop codon ochre (TAA), was found by comparison with two Pinus species, thus creating two additional potential stop codons.